Atopic dermatitis treatment method

ABSTRACT

Compositions are provided for treating atopic dermatitis, other atopic diseases and other inflammatory or allergic skin disorders. The compositions include proteins from Molluscum Contagiosum Virus (MCV), or fragments, variants, analogs, and derivatives thereof which exhibit AD inhibiting activity. Examples of MCV proteins which exhibit AD inhibiting activity include MC148P1, MC148P2, MC148P3, other MC148P type proteins, and fragments, variants, analogs, and derivatives of MC148P1, MC148P2, MC148P3, and other MC148P type proteins which possess AD inhibiting activity. The fragments, variants, analogs and derivatives may be less than 100% homologous to MCV proteins so long as they are sufficiently homologous such that AD inhibiting activity is preserved.

CROSS-REFERENCE

This application is a continuation of U.S. application Ser. No.09/920,897, filed Aug. 1, 2001, which is a continuation of U.S.application Ser. No. 09/624,748, filed Jul. 24, 2000, which is acontinuation of U.S. application Ser. No. 09/426,093, filed Oct. 22,1999, which are incorporated herein by reference in their entirety.

BACKGROUND OF THE INVENTION

1. Field of the Invention

The present invention relates to the treatment of inflammatory and/orallergic skin disorders and more particularly to the treatment of AtopicDermatitis using compositions which include a protein derived fromMolluscum Contagiosum Virus (MCV).

2. Description of Related Art

Atopic Dermatitis (AD) is a genetically determined, reaginically (IgE)associated, chronic disease of the skin affecting approximately 8million adults and children in the United States. In AD, the skin isdry, easily irritated, subject to immediate hypersensitivity type ofallergic responses, typically scaly, often thickened, commonly red,frequently infected, sometimes exudative and above all itchy. Amongadults with AD, coexisting respiratory allergy (allergic rhinitis and/orasthma), has been reported to range from 63% to 85%.

Reaginic diseases, i.e. atopic diseases, are characterized by thecapacity to form IgE antibodies, on a genetic basis, resulting inimmediate hypersensitivity reactions upon exposure to many specificallergens, most prominent among which may be the house dust mite(Dermatophagoides pteronyssinus), but which also include pollens, moldsand danders. Multiple genetic factors contribute to expression of thisphenotype. Atopic susceptibility genes include those making severalmajor histocompatibility complex (HLA) class II molecules, IL-4 receptorproteins and IgE high affinity receptor proteins. An atopysusceptibility gene recently identified involves a guanine for adeninesubstitution at nucleotide 1902 of the IL-4 receptor gene, synthesizingthe α subunit of the IL-4 receptor on the surface of B lymphocytes whichresults in an arginine for glutamine substitution at peptide position576 (R576). This substitution was found in 57% of patients with AD, butin only 17% of non-atopic controls, p=0.001. A majority of subjectsidentified as carrying a single copy of the mutant allele had atopy,suggesting a dominant effect, yet penetrance is modified by otherfactors, since others carrying allele R576 lacked atopy. R576 alters thebinding profile of the adjacent phosphorylated tyrosine residue, whichimpairs enzyme mediated termination of signaling of cytokine receptors,causing sustained or exaggerated receptor signaling. (NEJM 337:172 5,1997).

The frequent chronic infections that occur on the skin of AD patientsappears to result both from the defective barrier of AD skin and from animpaired immune response, e.g. upon testing with trichopyton antigen,patients with AD show immediate rather than the normal delayed immuneresponse. The most common of the microbes infecting AD skin isStaphylococcus aureus (Staph). In AD skin, Staph induces and exacerbatesitching, increases inflammation and provokes oozing and eczematization.Of those who ooze, 100% will culture out Staph. Of those who don't, themajority will still culture out Staph, though less massively. About 50%of patients with AD produce IgE directed against Staph toxins. Viruses,too, more readily grow in AD skin than in normal skin, including herpes,wart and molluscum viruses.

There is no known cure for AD. The many treatment approaches attest theinadequacy and limitations of each. In briefest outline, thesetreatments include avoidance of soap and water, hydrating the skin,dietary restrictions, avoidance of irritants and allergens in theenvironment, tars, antihistamines, hyposensitization, corticosteroids,antibacterials, antifungals, ultraviolet light, leukotriene blockers,inhibitors of mast cell content release, evening primrose oil, Chineseherbal teas, pentoxifylline, azathioprine, cyclosporin A,cyclophosphamide, tacrolimus, interferon γ, thymopentin andphosphodiesterase inhibitors. The corticosteroids are most commonly usedin clinical practice, but suffer from incomplete responses,tachyphylaxis, induction of atrophy and the potential of suppression ofthe pituitary-adrenal axis if used widely enough, long enough andpotently enough.

The invention also relates to a kit which includes a compositionaccording to the present invention. The kit may optionally includemultiple separately packaged portions of the composition, where eachportion is in an amount suitable for a single administration or formultiple administrations, e.g. administration from a tube or a jar. Thekit may also optionally include instructions regarding theadministration of the composition to a patient having AD, other atopicdiseases and other inflammatory or allergic skin disorders. In one ofmany variations, the instructions may teach how locally to administerthe composition to the patient.

SUMMARY OF THE INVENTION

The invention relates to compositions for treating atopic dermatitis(AD), other atopic diseases (including asthma, allergic rhinitis, hives)and other inflammatory and/or allergic disorders of the skin. Thecompositions according to the present invention include proteins fromMolluscum Contagiosum Virus (MCV), or fragments, variants, analogs, andderivatives thereof which exhibit AD inhibiting activity. Examples ofMCV proteins which exhibit AD inhibiting activity include MC148P1,MC148P2, MC148P3, other MC148P type proteins, and fragments, variants,analogs, and derivatives of MC148P1, MC148P2, MC148P3, and other MC148Ptype proteins which possess AD inhibiting activity. The fragments,variants, analogs and derivatives may be less than 100% homologous toMC148P1, MC148P2, MC184P3 so long as they are sufficiently homologoussuch that AD inhibiting activity is preserved. Collectively, the aboveMCV proteins, fragments, variants, analogs and derivatives are referredto herein as MC148 proteins (MC148P).

In one embodiment, the composition is suitable for topical applicationto a portion of patient's skin which exhibits AD signs and/or symptoms.In another embodiment, the composition is adapted for delivery by otherroutes including by injection intravenously, intramuscularly,subcutaneously or intradermally or by electroporation or iontophoresis.The composition may be delivered systemically or it may be deliveredremotely or locally at or near a portion of patient's skin whichexhibits AD signs and/or symptoms.

The invention also relates to a method for treating AD as well as otheratopic diseases and other inflammatory or allergic skin disorders (seeabove) which includes administering a composition according to thepresent invention to patients with AD, other atopic diseases and otherinflammatory or allergic skin disorders.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1A illustrates the DNA sequence (SEQ. ID. No. 1) of MCV type 1, ORF148 R.

FIG. 1B illustrates the amino acid sequence (SEQ. ID. No. 2) of theprotein produced from the DNA sequence of MCV type 1, ORF 148 R,illustrated in FIG. 1A.

FIG. 2A illustrates the DNA sequence (SEQ. ID. No. 3) of MCV type 2, ORF148 R.

FIG. 2B illustrates the amino acid sequence of the protein produced fromthe DNA sequence (SEQ. ID. No. 4) of MCV type 2, ORF 148 R, illustratedin FIG. 2A.

FIG. 3 illustrates the DNA sequence (SEQ. ID. No. 5) of MCV ORF 148 Rfrom the index case—the DNA of which is identical to that of MCV type 1,ORF 148 R, shown for nucleotides 20 to 312.

FIG. 4A illustrates the clinically demonstrable inhibitory effect of MCVupon a field of AD at a focal distance of 6 inches.

FIG. 4B illustrates the clinically demonstrable inhibitory effect of MCVupon field of AD at a focal distance of 2 inches.

FIG. 5A illustrates the microscopically demonstrable inhibitory effectof MCV upon a field of AD.

FIG. 5B illustrates the appearance of AD in the same patient shown inFIG. 5A at the same time at a site of AD remote from MCV.

DETAILED DESCRIPTION OF THE INVENTION

The compositions of the present invention include MC148 proteins whichexhibit AD inhibiting activity. As noted above, these proteins may beMCV proteins which exhibit AD inhibiting activity such as MC148P1,MC148P2, MC148P3, as well as fragments, variants, analogs, andderivatives of MCV proteins which exhibit AD inhibiting activity. Thefragments, variants, analogs and derivatives may be less than 100%homologous to an MCV protein so long as they are sufficiently homologoussuch that AD inhibiting activity is preserved.

Molluscum Contagiosum Virus (MCV) is a large DNA virus of the Poxfamily. MCV causes small, harmless lesions in the skin of infectedpersons. These small bumps (papules) resemble pimples that typicallyappear domed and shiny and that usually show a small central pit. MCVcan be spread from person to person by direct skin contact and byfomites. It is harmless and possesses no cancerous potential.

MC148P1 is natively produced by MCV type 1. MC148P2 is natively producedby MCV type 2.

The Applicant has observed in his clinical practice of medicine theinhibitory effect of MCV upon AD. The inhibitory effect endured at least3 years, i.e. for the duration of the MCV infection, any clinical sideeffect and without tachyphylaxis. By viewing micrographs at focaldistances of 6 inches (FIG. 4A} and of 2 inches (FIG. 4B) one sees afield of AD manifest as mildly scaly, somewhat lichenified reddish brownskin. Clear zones of clinically normal skin surround each papule of MCV.The zone of inhibition around each MCV papule may be viewed as analogousto the zone of inhibition around a penicillin disk on an agar platestreaked with Streptococci. The therapeutic implications are alsoanalogous.

FIG. 5A shows a photomicrograph of a biopsy taken at the edge of andimmediately adjacent to a papule of MCV. The edge of the papule is seenon the lower left of the panel. The top of the panel shows the base ofthe epidermis. A paucity of mononuclear cells is seen around small bloodvessels of the superficial dermal plexus. The lack of inflammation inthe dermis adjacent to the MCV papule resembles the appearance of thedermis in normal, i.e. non-AD skin. By contrast, FIG. 5B shows aphotomicrograph of a biopsy taken concurrently from a similar area of ADon the same patient but remote from any MCV papule. The moderatelydense, predominantly lymphohistocytic infiltrate admixed witheosinophils is seen not only around the blood vessels of the superficialplexus, but also around vessels of the papillae and those of the upperreticular dermis. The inflammatory cells extend into the interstitium.

Applicant interprets the above observations to show that MCV produces aprotein that inhibits the signs and symptoms of AD. This protein isbelieved to be MC148P1. Published work indicates that MC148P2 and otherMC148 proteins to share the same or similar anti-inflammatoryproperties. (Krathwohl et al. in Proc Nat'l Acad Sci 94: 9875-9880;Buget J. J. in Virology 242: 51-59, 1998; Damon, I. et al. in Proc Nat'lAcad Sci 95:6403-6407, 1998.)

Standard type 1 MCV is the major type of MCV found in nature andconsists of 190,289 base pairs. This comprises the entire genome,excepting covalently closed terminal hairpin loops. This genome wasdeposited in Gen Bank (accession number U60315) as described inSenkevich et al. “Genome Sequence of a Human Tumorigenic Poxvirus:Prediction of Specific Host Response-Evasion Genes”, Science273:813-816, 1996. MCV type 1 includes a DNA sequence of 312 bases,identified as ORF 148R, that encode a protein 104 amino acids in lengthreferred to herein as MC148P1. Synthesis, characterization and effectsof MC148P (MC148P1) are discussed in Damon et al. “Broad SpectrumChemokine Antagonistic Activity of a Human Poxvirus Chemokine Homolog”,Proc. Nat'l. Acad. Sci. USA, 95:6403-6407, 1998 and in Krathwohl et al“Functional Characterization of the C—C Chemokine-like Molecules Encodedby Molluscum Contagiosum Virus Types 1 and 2”, Proc. Natl. Acad. Sci.USA 94:9875-9880, 1997.

The DNA sequence of MCV148 type 1 (SEQ ID NO:1) is illustrated in FIG.1A and the amino acid sequence of MC148P1 (denoted as MC 148P by Damonet al and as MC 148R1 Protein by Krathwohl et al) (SEQ ID NO:2) isprovided in FIG. 1B.

MC 148P2 (also denoted MC148R 2 Protein), produced by MCV type 2, is avariant of MC148P1. MC148P2 is also 104 amino acids in length. The DNAsequence for MCV type 2 (SEQ ID NO:3) is illustrated in FIG. 2A and theamino acid sequence of MC148P2 (SEQ ID NO:4) is illustrated in FIG. 2B.The DNA sequence for MC148R2 has been deposited in Gen Bank (Accessionnumber U96749) by Krathwohl et al, as referenced above.

The MC148R2 has 89% homology to MC148R1. Amino acid sequences of MC148R2Protein (MC148P2) showed 87% homology with those of MC148R1 Protein(MC148P1) for complete sequences and 86% homology when the putativeleader sequence was removed. From the NH2 terminus, the leader sequencesof MC148P1 and MC148P2 consist of 24 amino acids of which 20 amino acidsshare identical positions. The chemokine activation domain, foundbetween positions 24 and 25 of MC148P1 and MC148P2, is absent in both.The 5 amino acids of positions 25-29, comprising the hypotheticalreceptor binding site, are identical except at position 26 where MC148P2bears a serine substitution for the alanine residue found in mostisolates of MC148P1. This substitution at position 26 does not appear toaffect the inhibitory activity of either type of MC148P. The leucine atposition 47 from the amino terminus, correlated with the ability ofMC148P to inhibit neutrophil chemotaxis, is conserved in MC148P1 andMC148P2.

Further, the amino acid sequences of MC148P1 and MC148P2 sharesignificant homology with CC (β) chemokines such as MacrophageInflammatory Protein-1α (MIP-1α) (Krathwohl et al, see above) and CC (β)chemokines including MIP-1α, RANTES, Macrophage Chemotactic Proteins-1and -3 (MCP-1 and MCP-3) (Damon et al, see above). The amino acidsequences of MC148P1 and of MC148P2 also share significant homology withCXC (α) chemokines SDF-1 for the attraction of monocytes and lymphocytesand IL-8 for the attraction of neutrophils. MC148P1 and MC148P2 sharethe identical positions of the 4 canonical cysteine residues with theabove mentioned CC and the CXC chemokines at positions 30, 31, 59 and 75of the respective amino acid chains. Taken together, these structuralhomologies may best account for the capacity of MC148P to inhibit thechemotaxis of human peripheral blood mononuclear cells (Krathwohl et al)and of monocytes, lymphocytes and neutrophils (Damon et al). Theinhibition results from the direct binding of MC148P to chemokinereceptors (Damon et al).

1. Compositions According to the Present Invention

The invention relates to compositions adapted for the treatment ofAtopic Dermatitis (AD), other atopic diseases and other inflammatory orallergic skin disorders. These compositions comprise a protein orsequence of amino acids selected from the group consisting of: MC148P1,MC148P2, MC148P3, another MC148P type protein, and a fragment, variant,analog, or derivative of these proteins which possesses AD inhibitingactivity.

A. Fragments of MC148P

Fragments of MC148P may be any amino acid sequence which is sufficientlyhomologous to a MC148P that AD inhibiting activity is preserved. Thesefragments may be generated by genetic engineering of translation stopsites within the coding region (discussed below). Alternatively,treatment of the MC148P with proteolytic enzymes, known as proteases,can produce a variety of N-terminal, C-terminal and internal fragments.Examples of fragments may include contiguous portions of an MC148P of 6,7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25,30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, or moreamino acids in length. These fragments may have primary, secondary(β-sheets, α-sheets, or other), tertiary and quaternary structures,including domains and loops.

These fragments may be purified according to known methods, such asprecipitation (e.g. ammonium sulfate), HPLC, ion exchangechromatography, affinity chromatography (including immunoaffinitychromatography) or various size separations (sedimentation, gelelectrophoresis, gel filtration).

B. Variants of MC148P

Variants of MC148P for inclusion in the compositions of the presentinvention can be substitutional, insertional or deletion variants ofMC148P. Deletion variants lack one or more residues of the nativeprotein which are not essential for function or immunogenic activity,and are exemplified by the variants lacking a leader sequence describedabove. Another common type of deletion variant is one lacking secretorysignal sequences or signal sequences directing a protein to bind to aparticular part of a cell. Insertional mutants typically involve theaddition of material at a non-terminal point in the polypeptide. Thismay include the insertion of an immunoreactive epitope or simply asingle residue. Terminal additions, called fusion proteins, arediscussed below.

Substitutional variants typically contain the exchange of one amino acidfor another at one or more sites within the protein, and may be designedto modulate one or more properties of the polypeptide, such as stabilityagainst proteolytic cleavage, without loss of other functions orproperties. Substitutions of this kind preferably are conservative, thatis, one amino acid is replaced with another of similar shape and charge.Conservative substitutions are well known in the art and include, forexample, the changes of: alanine to serine; arginine to lysine;asparagine to glutamine or histidine; aspartate to glutamate; cysteineor threonine to serine; glutamate to aspartate; glycine or leucine toproline; histidine to asparagine, lysine or glutamine; isoleucine toleucine or valine; leucine to valine; lysine to arginine; tyrosine tophenylalanine or tryptophan; the reverse of the above changes; othersubstitutions.

The following is a discussion based upon changing of the amino acids ofa protein to create an equivalent, or even an improved,second-generation molecule. For example, certain amino acids may besubstituted for other amino acids in a protein structure withoutappreciable loss of interactive binding regions of antibodies or bindingsites on substrate molecules. Since it is the interactive capacity andnature of a protein that defines that protein's biological functionalactivity, certain amino acid substitutions can be made in a proteinsequence, and its underlying DNA coding sequence, and neverthelessobtain a protein with like properties. It is thus contemplated by theinventors that various changes may be made in the DNA sequences of geneswithout appreciable loss of their biological utility or activity, asdiscussed below.

In making such changes, the hydropathic index of amino acids may beconsidered. The importance of the hydropathic amino acid index inconferring interactive biologic function on a protein is generallyunderstood in the art (Kyte & Doolittle, 1982). It is accepted that therelative hydropathic character of the amino acid contributes to thesecondary structure of the resultant protein, which in turn defines theinteraction of the protein with other molecules, for example, enzymes,substrates, receptors, DNA, antibodies, antigens, and the like.

Each amino acid has been assigned a hydropathic index on the basis oftheir hydrophobicity and charge characteristics (Kyte& Doolittle, 1982),these are: isoleucine (+4.5); valine (+4.2); leucine (+3.8);phenlyalanine (+2.8); cysteine/cystine (+2.5); methionine (+1.9);alanine (+1.8); glycine (−0.4); threonine (−0.7); serine (−0.8);tryptophan (−0.9); tyrosine (−1.3); proline (−1.6); histidine (−3.2);glutamate (−3.5); glutamine (−3.5); aspartate (−3.5); asparagine (−3.5);lysine (−3.9); and arginine (−4.5).

Certain amino acids may be substituted by other amino acids having asimilar hydropathic index or score and still result in a protein withsimilar biological activity, i.e. still obtain a biological functionallyequivalent protein. In making such changes, the substitution of aminoacids whose hydropathic indices are within +/−2 is preferred, thosewithin are within +/−1 are particularly preferred, and those within+/−0.5 even more particularly preferred.

The substitution of like amino acids can be made effectively on thebasis of hydrophilicity. U.S. Pat. No. 4,554,101, incorporated herein byreference, states that the greatest local average hydrophilicity of aprotein, as governed by the hydrophilicity of its adjacent amino acids,correlates with a biological property of the protein. As detailed inU.S. Pat. No. 4,554,101, the following hydrophilicity values have beenassigned to amino acid residues: arginine (+3.0); lysine (+3.0);aspartate (+3.0+/−1); glutamate (+3.0+/−1); serine (+0.3); asparagine(+0.2); glutamine (+0.2); glycine (0); threonine (−0.4); proline(−0.5+1-1); alanine (−0.5); histidine (−0.5); cysteine (−1.0);methionine (−1.3); valine (−1.5); leucine (−1.8); isoleucine (−1.8);tyrosine (−2.3); phenylalanine (−2.5); tryptophan (−3.4).

It is understood that an amino acid can be substituted for anotherhaving a similar hydrophilicity value and still obtain a biologicallyequivalent and immunologically equivalent protein. In such changes, thesubstitution of amino acids whose hydrophilicity values are within +/−2is preferred, those that are within +/−1 are particularly preferred, andthose within +/−0.5 are even more particularly preferred.

As outlined above, amino acid substitutions are generally based on therelative similarity of the amino acid side-chain substituents, forexample, their hydrophobicity, hydrophilicity, charge, size, and thelike. Exemplary substitutions that take various of the foregoingcharacteristics into consideration are well known to those of skill inthe art and include: arginine and lysine; glutamate and aspartate;serine and threonine; glutamine and asparagine; and valine, leucine andisoleucine.

Another embodiment for the preparation of polypeptides according to theinvention is the use of peptide mimetics. Mimetics arepeptide-containing molecules that mimic elements of protein secondarystructure. See, for example, Johnson et al., “Peptide Turn Mimetics” inBIOTECHNOLOGY AND PHARMACY, Pezzuto et al. Eds., Chapman and Hall, NewYork (1993). The underlying rationale behind the use of peptide mimeticsis that the peptide backbone of proteins exists chiefly to orient aminoacid side chains in such a way as to facilitate molecular interactions,such as those of antibody and antigen. A peptide mimetic is expected topermit molecular interactions similar to the natural molecule. Theseprinciples may be used, in conjunction with the principles outlinedabove, to engineer second generation molecules having many of thenatural properties of MCV-type 1, type 2 and other type viral proteins,but with altered and even improved characteristics.

A specialized kind of insertional variant is the fusion protein. Thismolecule generally has all or a substantial portion of the nativemolecule, linked at the N- or C-terminus, to all or a portion of asecond polypeptide.

For example, fusions typically employ leader sequences from otherspecies to permit the recombinant expression of a protein in aheterologous host. Another useful fusion includes the addition of animmunologically active domain, such as an antibody epitope, tofacilitate purification of the fusion protein. Inclusion of a cleavagesite at or near the fusion junction will facilitate removal of theextraneous polypeptide after purification. Other useful fusions includelinking of functional domains, such as active sites from enzymes,glycosylation domains, cellular targeting signals or transmembraneregions.

The above proteins, fragments and/or variants for use in thecompositions of the present invention can be produced by recombinantexpression means or, if small enough, generated by an automated peptidesynthesizer.

C. Administration to a Patient

Compositions according to the present invention, for the treatment ofAD, other atopic diseases and other inflammatory or allergic skindisorders may include one or more pharmaceutically acceptable carriersto provide a pharmaceutically acceptable composition for delivery to apatient. The phrase “pharmaceutically or pharmacologically acceptable”refers to molecular entities and compositions that are unlikely toproduce adverse, allergic or other untoward reactions when administeredto an animal or a human patient. As used herein, “pharmaceuticallyacceptable carrier” includes any and all solvents, suspensions,dispersion media, coatings, oils antibacterial and antifungal agents,preservatives, detergents, emollients, astringents, ointments, creams,lotions, gels; occlusion techniques; iontophoresis, electroporation orother devices, isotonic and absorption delaying agents and the like,including the types of carriers referenced in Smith E W & Maibach H I:“Percutaneous Penetration Enhancers”, CRC Press, 1995, N. W. Boca Raton,Fla. Methods contemplated for delivery of the composition are notlimited to chemical penetration enhances, but also include non-chemicalmethods such as iontophoresis and electroporation. Preferredpharmaceutically acceptable carriers include sulfoxides such asdecylmethylsulfoxide, pyrrolidones and combinations of these (Azone),macromolecular microspheres and liposomes. Supplementary activeingredients also can be incorporated into the compositions. The use ofsuch carriers and penetration enhancers for pharmaceutically activeingredients is well known in the art.

The pharmaceutically acceptable compositions of the present inventionmay include any classical or nonclassical pharmaceutical preparationwhich includes an MC 148P or a fragment, variant, analog or derivativeof a MC 148P as an active ingredient. Administration of thesecompositions according to the present invention will be via any commonroute so long as the target tissue is available via that route. Becausethe treatment of skin disease is contemplated, the route is preferablyadapted for the treatment of skin, e.g. a localized delivery, optionallytopically or via subcutaneous injection. Iontophoresis, electroporationor other techniques can be used for administration of thepharmaceutically acceptable composition.

When the composition is delivered topically, the composition ispreferably applied directly to the area affected by the skin disease.For subcutaneously or other administration, the most desirable point ofdelivery need not necessarily be at or near the area affected by theskin disease.

The pharmaceutically acceptable compositions according to the presentinvention may include sterile aqueous solutions or dispersions orsuspensions for the extemporaneous preparation of sterile injectablesolutions or dispersions. The form should be sterile and should be fluidin embodiments where the fluid is to be delivered by injection. The formshould be stable under the conditions of manufacture and storage andshould be preserved against the contaminating action of microorganisms,such as bacteria and fungi. The carrier can be a solvent or dispersionmedium containing, e.g. water, ethanol, polyol (such as glycerol,propylene glycol, liquid polyethylene glycol and the like), suitablemixtures thereof, and vegetable or other oils. Proper fluidity can bemaintained, by way of example, by the use of a coating, such aslecithin, by the maintenance of the required particle size in the caseof dispersion and by the use of surfactants. The prevention of theaction of microorganisms can be brought about by various antibacterialand antifungal agents, e.g. parabens, chlorobutanol, phenol, sorbicacid, thimerosal, benzalkonium and the like. In many cases, it may bepreferable to include isotonic agents, e.g. sugars or sodium chloride.Prolonged absorption of the injectable compositions can be brought aboutby the use in the compositions of agents delaying absorption, e.g.aluminum monostearate and gelatin.

Sterile injectable solutions may be prepared by incorporating the activeingredients in the required amount in the appropriate solvent withvarious of the other ingredients enumerated above, as required, followedby filtered sterilization. Generally, dispersions are prepared byincorporating the various sterilized active ingredients into a sterilevehicle which contains the basic dispersion medium and the requiredother ingredients from those enumerated above. In the case of sterilepowders for the preparation of sterile injectable solutions, thepreferred methods of preparation are vacuum-drying and freeze-dryingtechniques which yield a powder of the active ingredient plus anyadditional desired ingredient from a previously sterile-filteredsolution thereof.

The compositions of the present invention may be formulated in a neutralor salt form. Pharmaceutically acceptable salts include acid additionsalts (formed with the free amino groups of the protein) which aregenerally formed with inorganic acids such as, e.g. hydrochloric orphosphoric acids, or organic acids such as acetic, oxalic, tartaric,mandelic and the like. Salts formed with free carboxyl groups can alsobe derived from inorganic bases such as, e.g. sodium, potassium,ammonium, calcium, or ferric hydroxides, and such organic bases asisopropylamine, trimethylamine, histidine, procaine and the like.

Upon formulation, solutions may be administered in a manner compatiblewith the dosage formulation and in such amount as is therapeuticallyeffective. The formulations are easily administered in a variety ofdosage forms such as injectable solutions, and the like. For parenteraladministration in an aqueous solution, for example, the solution shouldbe suitably buffered if necessary and the liquid diluent first renderedisotonic with sufficient saline or glucose. In this connection, sterileaqueous media which can be employed will be known to those of skill inthe art in light of the present disclosure. For example, one dosagecould be dissolved in 1 ml of isotonic NaCl solution and either added to1000 ml of hypodermo-clysis fluid or injected at the proposed site ofinfusion (see for example, “Remington's Pharmaceutical Sciences” 15thEdition, pages 1035-1038 and 1570-1580). Some variation in dosage willnecessarily occur depending on the condition of the subject beingtreated. The person responsible for administration will, in any event,determine the appropriate dose for the individual subject. Moreover, forhuman administration, preparations should meet sterility, pyrogenicity,general safety and purity standards as required by FDA Office ofBiologics Standards.

2. Descriptions of FIGS. 4A-5B

FIGS. 4A-4B illustrate the clinically demonstrable inhibitory effect ofMCV upon a field of AD.

In FIGS. 4A and 4B the patient (PR) is a light skinned African Americanman, 16 years of age at the time these photos were taken. PR has had achronic, widespread, waxing and waning atopic dermatitis. He has carrieda persistent and heavy infection of Molluscum contagiosum virus (MCV).The numerous light reddish tan, mildly lucent, sometimes umbilicatedhillocks shown on the clinical photos are papules of epidermis infectedwith MCV. Many of the papules are 5 mm in diameter (range 3 to 7 mm).

FIG. 4A photograph was taken at a focal distance of 6 inches and FIG. 4Bphotograph at 2 inches. FIG. 4A shows a background of atopic dermatitismanifest as mildly scaly, somewhat lichenified, reddish brown skin.Clear zones of clinically normal skin surround each papule of MCV. Theclear zones range from 3 to 8 mm in dimension from the edge of a MCVpapule to the edge of the background dermatitis. There appears to be arough correlation between the size of a MCV papule and the width of theclear zone around it. The lack of a direct linear correlation could beassociated with colonization of the skin with Staphylococcus aureus. S.aureus frequently colonizes the skin of patients with atopic dermatitisand almost invariably exacerbates the dermatitis. PR has proven S.aureus skin infections.

FIG. 4B includes a photograph is taken of an area of more severe atopicdermatitis characterized by a dark grey brown, markedly lichenified skinwith prominent scale. Four frank excorations are demonstrable. (Personswith atopic dermatitis almost always itch). In the center of thephotograph are 2 MCV papules around which are clear zones of normalskin. Hence the anti-inflammatory effect to MCV is sufficiently powerfulto suppress even severely dermatitic atopic skin.

The zone of inhibition around each MCV papule may be viewed as analogousto the zone of inhibition around a penicillin disk on an agar platestreaked with Streptococci. The therapeutic implications are alsoanalogous.

FIGS. 5A-5B illustrates the microscopically demonstrable inhibitoryeffect of MCV upon a field of AD.

The photomicrograft of the biopsy depicted in FIG. 5A was taken from anarea of atopic dermatitis on the skin of PR immediately adjacent to apapule of Molluscum contagiosum virus (MCV). The edge of the papule isseen on the lower left, and within the lower spinous cell layer of thepapule, the cytoplasm of the keratinocytes contains Molluscum bodies,visible on light microscopy as deposits of eosinophilic amorphousmaterial. The top of the photomicrograph shows the base of the adjacentepidermis. Moderate numbers of fibroblasts are found in a modifiedconnective tissue around the Molluscum papule. A paucity of mononuclearcells is seen around small blood vessels of the superficial dermalplexus. There is a mild to moderate acanthosis, seen here in the lowerpart of the epidermis. There is scant inflammatory infiltrate within theconnective tissue in the region of the Molluscum papule, extending farlaterally into the papillary and reticular parts of the dermis. The lackof inflammation in the dermis adjacent to the Molluscum papule resemblesthe milieu characteristic of normal skin. (H&E, 100).

The photomicrograft of the biopsy depicted in FIG. 5B was takenconcurrently (the same date and time) from similar area of atopicdermatitis on the skin of PR remote from the Molluscum papule shown inFIG. 5A. The top of the photomicrograph shows a markedly acanthoticepidermis with a central adherent crust denoting a site of excoriation,due to the incessant itching characteristic of atopic dermatitis. Theitching, in turn, is secondary to inflammatory mediators of diverseorigin—many of which are the products of the inflammatory cells whichinfiltrate the skin of patients with atopic dermatitis. The moderatelydense, predominantly lymphohistiocytic infiltrate is seen here not onlyaround the blood vessels of the superficial plexus, but also aroundvessels of the upper reticular dermis serving that plexus and aroundvessels of the papillae. The infiltrate of inflammatory cells extendsinto the interstitium. Lymphocytic exocytosis is also present. (H & E,100×). With further magnification, eosinophils can also be seen withinthe inflammatory infiltrate on this photomicrograph.

While the present invention is disclosed by reference to the preferredembodiments and examples detailed above, it is to be understood thatthese examples are intended in an illustrative rather than limitingsense, as it is contemplated that modifications and combinations willreadily occur to those skilled in the art, which modifications andcombinations will be within the spirit of the invention and the scope ofthe appended claims.

1. A method, comprising: administering to a patient having atopicdermatitis a composition comprising a MC148P protein which possessesatopic dermatitis inhibiting activity.
 2. The method according to claim1, wherein the MC148P protein is selected from the group consisting ofMC148P1, MC148P2, MC148P3 and fragments, variants, analogs, andderivatives of MC148P1, MC148P2, and MC148P3 which possess atopicdermatitis inhibiting activity.
 3. The method according to claim 1,wherein administering the composition includes topically applying thecomposition.
 4. The method according to claim 1, wherein administeringthe composition includes injecting the composition.
 5. The methodaccording to claim 1, wherein administering the composition includesiontophoresis.
 6. The method according to claim 1, wherein administeringthe composition includes electroporation.
 7. The method according toclaim 1, wherein the composition includes a dimethyl sulfoxide carrier.8. The method according to claim 1, wherein the composition includes anazone carrier.
 9. The method according to claim 1, wherein thecomposition includes a liposomal carrier.
 10. The method according toclaim 1, wherein the composition is delivered locally to an area whereatopic dermatitis is believed to be present.
 11. The method according toclaim 1 the composition is delivered to a patient with an atonic diseaseor allergic skin disorder
 12. A kit, comprising: multiple separatelypackaged portions of a composition adapted for treating atopicdermatitis comprising a MC148P protein which possesses atopic dermatitisinhibiting activity.
 13. The kit according to claim 12, wherein theMC148P protein is selected from the group consisting of MC148P1,MC148P2, MC148P3 and fragments, variants, analogs, and derivatives ofMC148P1, MC148P2, and MC148P3 which possess atopic dermatitis inhibitingactivity.
 14. The kit according to claim 12, wherein the kit furtherincludes instructions teaching administration of the composition to apatient having atopic dermatitis.
 15. The kit according to claim 14,wherein the instructions teach locally delivering the composition to anarea adjacent where atopic dermatitis is believed to be present.
 16. Thekit according to claim 14, wherein the instructions teach topicallyapplying the composition.
 17. The kit according to claim 14, wherein theinstructions teach injecting the composition.
 18. The kit according toclaim 14, wherein the instructions teach administering the compositionthrough iontophoresis and/or electroporation.
 19. The kit according toclaim 12, wherein the composition includes a dimethyl sulfoxide carrier.20. The kit according to claim 12, wherein the composition includes anazone carrier.
 21. The kit according to claim 12, wherein the kit isalso used for the treatment of other atopic diseases and otherinflammatory and or allergic skin disorders.